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Marker composed of tissue snoRNA and kit used for predicting liver cancer recurrence risk

A marker and kit technology, applied in the field of detection kits for assessing the recurrence risk of hepatocellular carcinoma patients after undergoing resection, can solve the problems of difficulty in detection implementation, poor usability, and few of them, so as to avoid excessive treatment and achieve mature experimental methods. , the effect of simple detection process

Inactive Publication Date: 2017-01-04
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, although various models for predicting prognosis have emerged in an endless stream in recent years, there are still very few models that can be applied clinically.
The main problems are as follows: (1) The research results are not repeatable; Michiels reanalyzed the data of 7 published studies on tumor prognosis classifiers, and found that the results of 5 studies were not repeatable, and proposed training methods for classifier construction. The sample size of the group should be large enough, and it should be verified independently and multiple times to ensure the reproducibility of the results
(2) The classifier contains too much redundant information, and the usability is poor; the increase in the number of indicators can improve the prediction accuracy of the classifier. For example, the classifier built by the Hoshida team contains 186 genes, but this will also cause difficulties in detection execution. The problem of high cost brings difficulties to clinical application

Method used

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  • Marker composed of tissue snoRNA and kit used for predicting liver cancer recurrence risk
  • Marker composed of tissue snoRNA and kit used for predicting liver cancer recurrence risk
  • Marker composed of tissue snoRNA and kit used for predicting liver cancer recurrence risk

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1: Collection and preparation of tumor tissue samples

[0033] The inventors collected tumor tissue samples from patients with liver cancer (HCC) who underwent liver cancer resection between January 2006 and November 2011. These populations met the following inclusion criteria, and according to the principle of gender and age matching, set liver cancer and its control samples. Inclusion criteria: (1) primary liver cancer, newly diagnosed and underwent radical surgical resection; (2) aged between 18 and 80 years old; (3) without extrahepatic metastasis at the time of diagnosis; (4) without liver cancer before operation Other malignant diseases, and no anti-cancer treatment before postoperative recurrence; (5) No symptoms of severe organ disorders after postoperative.

[0034] Training group: 174 cases of HCC tumor tissue specimens who underwent liver cancer resection between January 2006 and December 2009.

[0035] Validation group: 109 cases of HCC tumor tissu...

Embodiment 2

[0039] Embodiment 2: gene chip and its data analysis

[0040] The inventor selected 5 cases of liver cancer tissue and 3 cases of normal liver tissue (paratumor liver tissue of hepatic hemangioma) for gene chip screening. These specimens were obtained from patients undergoing radical resection of HCC or resection of hemangioma in 2005-2006, all were confirmed by pathology, and were frozen in liquid nitrogen immediately after resection.

[0041] The chip was produced by CapitalBio Corp, and a total of 281 snoRNA levels were detected. After calibration of the obtained raw data, the inventors used the Significant Analysis of Microarray (SAM) analysis method to select differential snoRNAs, and finally screened and obtained 28 candidate snoRNAs for subsequent verification: ACA3, U15a, U19, ACA21, ACA31, U31, U35B , ACA38, U35b, snR38c, U42B, U44, ACA52, U52, U53, U54, U58b, U60, ACA61, U70, U75, U78, U81, U106, HBII-142, HBII-296, HBII-202, HBII-420 .

Embodiment 3

[0042] Embodiment 3: Real-time fluorescent quantitative PCR detects the level of snoRNAs in the specimens of the training group

[0043] 1. Tissue RNA Extraction

[0044] The present invention adopts Trizol reagent to extract, and specific steps are as follows: (1) For the tissue preserved in RNALater or liquid nitrogen, add 1ml Trizol ratio lysis cell per 50mg tissue; (2) Transfer Trizol lysate to 1.5ml Rnase free EP Shake and mix in the tube, place at room temperature for 5 minutes, add 1 / 5 of the lysate volume of chloroform, shake and mix, centrifuge at 12,000 g at 4°C for 15 minutes; absorb the supernatant, add an equal volume of isopropanol, place at room temperature for 10 minutes, and store at 4°C, Centrifuge at 12,000 g for 30 min; (3) Discard the supernatant, wash the precipitate twice with 70% ethanol, pour off the supernatant carefully, add an appropriate amount of DEPC water to dissolve the RNA after the alcohol evaporates, and store at -80°C for later use.

[004...

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Abstract

Disclosed are a marker composed of tissue snoRNA and a kit used for predicting the liver cancer recurrence risk. The invention discloses a marker used for predicting the postoperative recurrence risk of a patient who suffers from liver cancer. The maker is composed of one or several of the following snoRNA nucleic acid molecules: U15a, U19, ACA21, ACA31, U42b, U52 and / or ACA61. The invention further discloses a multi-gene kit for evaluating the postoperative recurrence risk of the patient who suffers from liver cancer. The kit comprises an agent for detecting the level of the following snoRNA nucleic acid molecules including U15a, U19, ACA21, ACA31, U42b, U52 and / or ACA61 in the tumor tissue of a patient who suffers from liver cancer. The kit can reflect the postoperative recurrence risk of a patient who suffers from liver cancer, allows excessive treatment to a low-risk patient to be avoided, and warns a high-risk patient. The kit is convenient for a clinician to adopt a personalized recurrence prevention scheme in time.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a detection kit for evaluating the recurrence risk of patients with hepatocellular carcinoma after resection. Background technique [0002] Primary hepatocellular carcinoma (referred to as liver cancer) is a highly malignant tumor, accounting for more than 90% of primary liver cancer. According to the "Global Cancer Report 2014" published by the World Health Organization, the incidence and mortality of liver cancer ranked fifth and second among malignant tumors in 2012. China is an area with a high incidence of liver cancer, and both new cases and deaths account for about half of the global cases. Surgical resection is currently one of the most effective methods for the treatment of liver cancer. However, even for patients who undergo surgical resection, the postoperative recurrence rate is still high, and the overall prognosis is not ideal. Studies have shown that post...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6886C12Q2600/178C12Q2600/158C12Q2600/118
Inventor 庄诗美杨金娥杨晓静
Owner SUN YAT SEN UNIV
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